胎肺发育中相关活性物质的表达及其调控意义
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摘要
目的
胎肺的发育过程存在着精细而严格的调控机制，在肺外形演变及呼吸功能建立
的过程中，胎肺细胞受到多种活性物质的调控，但是至今关于这类活性物质与胎肺形
态发生及功能分化的直接关系以及各种相关活性物质之间的调控关系则了解不多。
本研究观察小鼠胎肺发育分化过程中表皮生长因子受体（EGFR）、转化生长因子β
（TGF-β_1、TGF-β_2、TGF-β_3）和甲状腺转录因子-1（TTF-1）在支气管上皮细胞和
肺泡上皮细胞中的表达特征，并进行定性和定量分析研究，探讨它们在调节胎肺发
育和分化过程中的生物活性作用，以及相互之间的调控意义。
方法
合格昆明小鼠10-19天胎肺组织，共10组，生后及成年鼠肺组织，共5组，新鲜
取材；常规固定和包埋，切片5μm；兔疫组化SP法和ABC法检测EGFR、TGF-β_1、TGF-
β_2、TGF-β_3和TTF-1在肺组织中的表达特征；采用TIGER图象分析系统，对TTF-1的
表达强度进行定量分析。
结果
1．胎肺发育早期，EGFR的表达主要定位于近端支气管上皮，集中在支气管的
分叉处；胎18天，EGFR阳性反应达到最高峰；肺泡期，EGFR阳性反应主要在肺泡上
皮细胞，支气管上皮细胞着色趋淡；出生后，支气管上皮细胞EGFR重新呈阳性表达，
一周之后渐减弱。
2．TGF-β_1、TGF-β_2、TGF-β_3三者的分布定位基本一致，随着胎肺发育，其
暨南大学2001年硕士研究生毕业论文 胎肺发育中相关活性物质的夫达及其调控意义
在支气管上皮细胞的表达逐渐增强，在同一支气管上呈现区域性分布的特点：到发
育末期支气管上皮表达减弱，却在支气管周围间质呈强阳性：出生后，支气管上皮
呈弱阳性，肺泡上皮则无表达。
3．整个肺发育过程中均可检测到TTF刁的表达。主要定位于正在形成呼吸道的
呼吸上皮细胞核内，末端支气管明显比近端者强；出生后，TTFd移位于＊型肺泡
上皮细胞，支气管上皮细胞反应减弱。
4．随着胎肺的发育，远侧支气管上皮的TTF＊阳性细胞平均光密度（AOD）值
逐渐增加，体积（VOL）值逐渐减小，虽体积分光密度（VI OD）值也渐减小，但减
速较VOL减速明显缓慢；表明TTF－1因子密度的增加，与＊型肺泡上皮细胞的功能相
适应。
结论
在胎肺 发育的不同时期，EGFR、TGF－pl、TGF－6 2、TGF－p 3和HF－1因子在上皮
细胞的表达定位有迁移，免疫组化反应强弱也有差异变化，提示它们在胎肺不同发育
阶段发挥不同的功能。EGFR和TTF习具有促进细胞发育分化的作用，而TGF－p因子的
作用正相反，是一种有选择的抑制作用，因此推测这类相关活性物质在肺发育过程
中不仅对支气管树的构筑，还对呼吸道上皮细胞和肺泡上皮的成熟分化有重要的调
节作用。胎肺的正常发育有赖于这些活性物质之间的对立统一协调作用的共同调控。
OBJECTIVE
The development of the fetal lung involves precise and strict regulating mechanism.
During the process of the morphogenesis and cell differentiation, embryonic lung cells
are influenced by kinds of polypeptides. But little information is available on how they
directIy influence lung morphogenesis and differentiation and on how they interact on in
the developing lung. In this work, distributions of the interrelated proteins: the
epidermal growth factor receptor(EGFR), transforming groWth factor-9l, 62, 63(TGF-p l,
pz, ps) and thyroid transcription factor- l (TTF-l ) in fetal and postnatal mouse Iungs were
determined, their important function properties in regulating lung morphogenesis,
differentiation and maturation were studied, and their reIationships between each other
were discussed.
METHODS
By breeding KUNMING mice, the Iung specimens of fetal (from l0 to l9 day of
gestation) and of postnatal (from neonate to adult mice) were obtained. The expression
of the polypeptides (EGFR, TGF-6l, TGF-62, TGF-63, TTF-l ) were examined by
immunohistochemistry. TIGER image analysiser was used in quantitatively studying.
RESULTS
l It is showed that EGFR localized in the airway surface epithelium during early
3
lung development and its reaction reached the peak on fetal age Day l8. In saccular and
alveolar period, the positive reactions were mainly detected in alveolar cells.
Bronchiolar epithelial cells expressed EGFR again after birth, and then gradually
became weaker.
2 The localization patterns in the fetal 1ung for TGF-0l, TGF-62, TGF-93 were
essentially identical. At the early period, regional expression were observed in the
bronchioles. they were slighter in late gestation, but mesenchymal cells expressed at
high levels. In postnatal lung, no TGF-Pl, 62, 63 immunoreactivity was seen in the
alveolar cells.
3. TTF-l were expressed throughout fetal lung development, prominent in the
nuclei of distal lung buds, whose staining was more intense than that of airway epithelial
ceIls. After birth, TTF-l was restricted to type II cells and TTF-1 became weaker in
epithelial cells of trachea.
4. The average optical density(AOD) of TTF-l in the distal epithelial cells
increased gradually during the lung development. Though their volume(VOL) and
volume integrated optical density(VIOD) all decreased, the fOrmer dropped more
quickly than the latter, which showed the increasing density of TTF-l and its identical
function with type II cells.
CONCLUSION
The positive reactions of EGFR, TGF-6l, TGF-p2, TGF-P3 and TTF-l were
respectively different during different stages, which suggests that their functions are
changing in individual developmental periods. TGF-6 inhibits lung development, but on
the contrary, EGFR and TTF-l not only stimulate the process of branching
morphogenesis. but aIso modulate epitheIial maturation and differentiation. Normal lung
development depends on the unity of two opposites.
胎肺的发育过程存在着精细而严格的调控机制，在肺外形演变及呼吸功能建立
的过程中，胎肺细胞受到多种活性物质的调控，但是至今关于这类活性物质与胎肺形
态发生及功能分化的直接关系以及各种相关活性物质之间的调控关系则了解不多。
本研究观察小鼠胎肺发育分化过程中表皮生长因子受体（EGFR）、转化生长因子β
（TGF-β_1、TGF-β_2、TGF-β_3）和甲状腺转录因子-1（TTF-1）在支气管上皮细胞和
肺泡上皮细胞中的表达特征，并进行定性和定量分析研究，探讨它们在调节胎肺发
育和分化过程中的生物活性作用，以及相互之间的调控意义。
方法
合格昆明小鼠10-19天胎肺组织，共10组，生后及成年鼠肺组织，共5组，新鲜
取材；常规固定和包埋，切片5μm；兔疫组化SP法和ABC法检测EGFR、TGF-β_1、TGF-
β_2、TGF-β_3和TTF-1在肺组织中的表达特征；采用TIGER图象分析系统，对TTF-1的
表达强度进行定量分析。
结果
1．胎肺发育早期，EGFR的表达主要定位于近端支气管上皮，集中在支气管的
分叉处；胎18天，EGFR阳性反应达到最高峰；肺泡期，EGFR阳性反应主要在肺泡上
皮细胞，支气管上皮细胞着色趋淡；出生后，支气管上皮细胞EGFR重新呈阳性表达，
一周之后渐减弱。
2．TGF-β_1、TGF-β_2、TGF-β_3三者的分布定位基本一致，随着胎肺发育，其
暨南大学2001年硕士研究生毕业论文 胎肺发育中相关活性物质的夫达及其调控意义
在支气管上皮细胞的表达逐渐增强，在同一支气管上呈现区域性分布的特点：到发
育末期支气管上皮表达减弱，却在支气管周围间质呈强阳性：出生后，支气管上皮
呈弱阳性，肺泡上皮则无表达。
3．整个肺发育过程中均可检测到TTF刁的表达。主要定位于正在形成呼吸道的
呼吸上皮细胞核内，末端支气管明显比近端者强；出生后，TTFd移位于＊型肺泡
上皮细胞，支气管上皮细胞反应减弱。
4．随着胎肺的发育，远侧支气管上皮的TTF＊阳性细胞平均光密度（AOD）值
逐渐增加，体积（VOL）值逐渐减小，虽体积分光密度（VI OD）值也渐减小，但减
速较VOL减速明显缓慢；表明TTF－1因子密度的增加，与＊型肺泡上皮细胞的功能相
适应。
结论
在胎肺 发育的不同时期，EGFR、TGF－pl、TGF－6 2、TGF－p 3和HF－1因子在上皮
细胞的表达定位有迁移，免疫组化反应强弱也有差异变化，提示它们在胎肺不同发育
阶段发挥不同的功能。EGFR和TTF习具有促进细胞发育分化的作用，而TGF－p因子的
作用正相反，是一种有选择的抑制作用，因此推测这类相关活性物质在肺发育过程
中不仅对支气管树的构筑，还对呼吸道上皮细胞和肺泡上皮的成熟分化有重要的调
节作用。胎肺的正常发育有赖于这些活性物质之间的对立统一协调作用的共同调控。
OBJECTIVE
The development of the fetal lung involves precise and strict regulating mechanism.
During the process of the morphogenesis and cell differentiation, embryonic lung cells
are influenced by kinds of polypeptides. But little information is available on how they
directIy influence lung morphogenesis and differentiation and on how they interact on in
the developing lung. In this work, distributions of the interrelated proteins: the
epidermal growth factor receptor(EGFR), transforming groWth factor-9l, 62, 63(TGF-p l,
pz, ps) and thyroid transcription factor- l (TTF-l ) in fetal and postnatal mouse Iungs were
determined, their important function properties in regulating lung morphogenesis,
differentiation and maturation were studied, and their reIationships between each other
were discussed.
METHODS
By breeding KUNMING mice, the Iung specimens of fetal (from l0 to l9 day of
gestation) and of postnatal (from neonate to adult mice) were obtained. The expression
of the polypeptides (EGFR, TGF-6l, TGF-62, TGF-63, TTF-l ) were examined by
immunohistochemistry. TIGER image analysiser was used in quantitatively studying.
RESULTS
l It is showed that EGFR localized in the airway surface epithelium during early
3
lung development and its reaction reached the peak on fetal age Day l8. In saccular and
alveolar period, the positive reactions were mainly detected in alveolar cells.
Bronchiolar epithelial cells expressed EGFR again after birth, and then gradually
became weaker.
2 The localization patterns in the fetal 1ung for TGF-0l, TGF-62, TGF-93 were
essentially identical. At the early period, regional expression were observed in the
bronchioles. they were slighter in late gestation, but mesenchymal cells expressed at
high levels. In postnatal lung, no TGF-Pl, 62, 63 immunoreactivity was seen in the
alveolar cells.
3. TTF-l were expressed throughout fetal lung development, prominent in the
nuclei of distal lung buds, whose staining was more intense than that of airway epithelial
ceIls. After birth, TTF-l was restricted to type II cells and TTF-1 became weaker in
epithelial cells of trachea.
4. The average optical density(AOD) of TTF-l in the distal epithelial cells
increased gradually during the lung development. Though their volume(VOL) and
volume integrated optical density(VIOD) all decreased, the fOrmer dropped more
quickly than the latter, which showed the increasing density of TTF-l and its identical
function with type II cells.
CONCLUSION
The positive reactions of EGFR, TGF-6l, TGF-p2, TGF-P3 and TTF-l were
respectively different during different stages, which suggests that their functions are
changing in individual developmental periods. TGF-6 inhibits lung development, but on
the contrary, EGFR and TTF-l not only stimulate the process of branching
morphogenesis. but aIso modulate epitheIial maturation and differentiation. Normal lung
development depends on the unity of two opposites.
引文
1．K. L. Moore著 何泽涌译．人体发生学(面向临床的胚胎学) 人民卫生出版社
2．成令忠主编 组织学与胚胎学(第四版) 人民卫生出版社
3. Ten Have Opbroek AA. Ultrastructural features of type Ⅱ alveolar epithelial cells in early embryonic mouse lung. Anat Rec 1988 Aug; 221(4): 846-53
4. Ten Have Opbroek AA. Ultrastructural characteristics of inclusion bodies of type Ⅱ cells in late embryonic mouse lung. Anat Embryol Berl 1990: 181(4): 317-23
5．岳少杰、罗自强综述 肺表面活性物质结合蛋白的生理功能 国外医学(呼吸系统分册)1995；15：79-82
6．陈政良 SP-A的分子生物学 国外医学(分子生物学分册) 1997：19：241-245
7．陈习中综述、罗自强审校 肺表面活性物质蛋白研究进展 国外医学(生理病理科学与临床分册)1996；16：33-35
8．范永琛 成人型呼吸窘迫综合征与肺泡表面活性物质 国外医学(儿科分册)1994；21(1)：7-11
9．高树生综述、陈竹钦审校 生长因子在胚胎发生和胎儿生长中的作用 国外医学(妇产科学分册)1996：23(1)：16-20
10. Brown PI, Lam R, Lakshmanan J, Fisher DA. Transforming growth factor alpha in developing rats. Am J Physiol 1990; 259(2 Pt 1): E256-260
11. Han VK, D'Ercole AJ, Lee DC. Expression of transforming growth factor alpha during development. Can J Physiol Pharmacol 1988; 66(8): 1113-1121
12．张平 转化生长因子-β族信号转导与肿瘤 中国病理生理杂志 2001：17(4)：377-380
13．周琪、谭景和 多肽生长因子与哺乳动物早期胚胎发育 生物技术1996；6(3)：1-7
14．蔡家新、童坦君 表皮生长因子对鼠胚成纤维细胞细胞周期的调节作用 生物化学杂志1994；10：218-222
15. Lonigtro R, De Felice M, Biffali E, Macchia PE, Damante G, Asteria C, Di Lauro R. Expression of thyroid transcription factor-1 gene can be regulated at the transcriptional and posttranscriptional levels. Cell Growth Differ 1996; 7(2): 251-61
16. Tell JG, Perrone L, Fabbro D, Pellizzari L, Pucillo C, De Felice M, Acquaviva R, Formisano S, Damante G. Structural and functional properties of the N transcriptional activation domain of thyroid transcription factor-1: similarities with the acidic activation domains. Biochem J 1998; 392(Pt2): 395-403
17．陈玲 表皮生长因子及其生物学效应 国外医学(儿科学分册)1997；24：240-242
18．江凌晓综述、封志纯审校 生长因子与胎肺发育 国外医学(生理病理科学与临床分册)1999；19(6)：489-492
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33. Wang X, Jin Q; Xu J, Shen W, Wang J, Post M. TGF-beta 3 inhibits the increased gene expressions of pulmonary surfactant proteins induced by
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38． Li J, Gao E, Mendelson CR. Cyclic AMP-responsive expression of the surfactant protein-A gene is mediated by increased DMA binding and transcriptional activity of thyroid transcription factor-1． J Biol Cjhem. 1998; 273(8) :4592-600
39． Kelly SE, Bachurski CJ, Burhans MS, Glasser SW. Transcription of the lung-specific surfactant protein C gene is mediated by thyroid transcription factor 1． J Biol Chem. 1996; 271(12) : 6881-6888
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42． Kumar AS, Venkatesh VC, Planer BC, Feinstein SI, Ballard PL. Phorbol ester down-regulation of lung surfactant protein B gene expression by cytoplasmic trapping of thyriod transcription factor-1 and hepatocyte nuclear factor-3． J Biol Chem 1997 Aug 15;272(33) :20764-20773
43． Hollenberg MD. Growth factors, their receptors and development. Am J-MedGen Et 1989;34 (1) :35-42
2．成令忠主编 组织学与胚胎学(第四版) 人民卫生出版社
3. Ten Have Opbroek AA. Ultrastructural features of type Ⅱ alveolar epithelial cells in early embryonic mouse lung. Anat Rec 1988 Aug; 221(4): 846-53
4. Ten Have Opbroek AA. Ultrastructural characteristics of inclusion bodies of type Ⅱ cells in late embryonic mouse lung. Anat Embryol Berl 1990: 181(4): 317-23
5．岳少杰、罗自强综述 肺表面活性物质结合蛋白的生理功能 国外医学(呼吸系统分册)1995；15：79-82
6．陈政良 SP-A的分子生物学 国外医学(分子生物学分册) 1997：19：241-245
7．陈习中综述、罗自强审校 肺表面活性物质蛋白研究进展 国外医学(生理病理科学与临床分册)1996；16：33-35
8．范永琛 成人型呼吸窘迫综合征与肺泡表面活性物质 国外医学(儿科分册)1994；21(1)：7-11
9．高树生综述、陈竹钦审校 生长因子在胚胎发生和胎儿生长中的作用 国外医学(妇产科学分册)1996：23(1)：16-20
10. Brown PI, Lam R, Lakshmanan J, Fisher DA. Transforming growth factor alpha in developing rats. Am J Physiol 1990; 259(2 Pt 1): E256-260
11. Han VK, D'Ercole AJ, Lee DC. Expression of transforming growth factor alpha during development. Can J Physiol Pharmacol 1988; 66(8): 1113-1121
12．张平 转化生长因子-β族信号转导与肿瘤 中国病理生理杂志 2001：17(4)：377-380
13．周琪、谭景和 多肽生长因子与哺乳动物早期胚胎发育 生物技术1996；6(3)：1-7
14．蔡家新、童坦君 表皮生长因子对鼠胚成纤维细胞细胞周期的调节作用 生物化学杂志1994；10：218-222
15. Lonigtro R, De Felice M, Biffali E, Macchia PE, Damante G, Asteria C, Di Lauro R. Expression of thyroid transcription factor-1 gene can be regulated at the transcriptional and posttranscriptional levels. Cell Growth Differ 1996; 7(2): 251-61
16. Tell JG, Perrone L, Fabbro D, Pellizzari L, Pucillo C, De Felice M, Acquaviva R, Formisano S, Damante G. Structural and functional properties of the N transcriptional activation domain of thyroid transcription factor-1: similarities with the acidic activation domains. Biochem J 1998; 392(Pt2): 395-403
17．陈玲 表皮生长因子及其生物学效应 国外医学(儿科学分册)1997；24：240-242
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